As an alternative to the present toxic chromate-based coating system now in use, the Mg-rich primer technology has been designed to protect Al alloys (in particular Al 2024 T3) and developed in analogy to Zn-rich primers for steel substrate. As an expansion of this concept, metal-rich primer systems based on Mg alloy particles as pigments were studied. Five different Mg alloy pigments, AM60, AZ91B, LNR91, AM503 and AZG, were characterized by using the same epoxy-polyamide polymer as binder, a same dispersion additive and the same solvent. Different Mg alloy-rich primers were formulated by varying the Mg alloy particles and their pigment volume concentrations (PVC). The electrochemical performance of each Mg alloy-rich primer after the cyclic exposure in Prohesion chamber was investigated by electrochemical impedance Spectroscopy (EIS). The results indicated that all the Mg alloy-rich primers could provide cathodic protection for AA 2024 T3 substrates. However, the Mg alloys as pigments in metal-rich primers seemed to exhibit the different anti-corrosion protection performances, such as the barrier properties, due to the different properties of these pigments. In these investigations, multiple samples of each system were studied and statistical methods were used in analyzing the EIS data. From these results, the recommendation for improved EIS data analysis was made. CPVC studies were carried out on the Mg alloy-rich primers by using three Mg alloy pigments, AM60, AZ91B and LNR91. A modified model for predicting CPVC is proposed, and the results showed much better agreement between the CPVC values obtained from the experimental and mathematical methods. Using the data from the AM60 alloy pigment system, an estimate of experimental coarseness was done on a coating system, the first time such an estimate has been performed. By combining various surface analysis techniques, such as scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS) and confocal Raman microscopy, the oxidation products formed after exposure were identified. It was found that variation of Al content in Mg alloy could significantly affect the pH of the microenvironment in the primer films and result in the formation of various oxidation products. / Air Force Office of Scientific Research (Grant No. 49620-02-1-0398)

As an alternative to the present toxic chromate-based coating system now in use, the Mg-rich primer technology has been designed to protect A1 alloys (in particular A1 2024 T3) and developed in analogy to Zn-rich primers for steel substrate. As an expansion of this concept, metal-rich primer systems based on Mg alloy particles as pigments were studied. Five different Mg alloy pigments. AM60, A719B, LNR91, AM503 and AZG, were characterized by using the same epoxy-polyamide polymer as binder, a same dispersion additive and the same solvent. Different Mg alloy-rich primers were formulated by varying the Mg alloy particles and their pigment volume concentrations (PVC). The electrochemical performance of each Mg alloy-rich primer alter the cyclic exposure in Prohesion chamber was investigated by electrochemical impedance Spectroscopy (EIS). The results indicated that all the Mg alloy-rich primers could provide cathodic protection for AA 2024 T3 substrates. However, the Mg alloys as pigments in metal-rich primers seemed to exhibit the different anti-corrosion protection performances, such as the barrier properties, due to the different properties of these pigments. In these investigations, multiple samples of each system were studied and statistical methods were used in analyzing the EIS data. From these results. the recommendation for improved EIS data analysis was made. CPVC studies were carried out on the Mg alloy-rich primers by using three Mg alloy pigments, AM60, A2918 and LNR91. A modified model for predicting CPVC is proposed, and the results showed much better agreement between the CPVC values obtained from the experimental and mathematical methods. Using the data from the AM60 alloy pigment system, an estimate of experimental coarseness was done on a coating system, the first time such an estimate has been performed. By combining various surface analysis techniques, such as scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS) and confocal Raman microscopy, the oxidation products formed alter exposure were identified. It was found that variation of A1 content in Mg alloy could significantly affect the pH of the microenvironment in the primer films and result in the formation of various oxidation products. / Air Force Office of Scientific Research (AFOSR) (Grant No. 49620-02-1-0398)

Indiana University-Purdue University Indianapolis (IUPUI) / BACKGROUND: Phosphoric acid treatment has been reported to be relevant for zirconia cleaning after contamination. However, its interaction with different primers is not well known and needs to be investigated. OBJECTIVES: To compare the shear bond strengths (SBS) of resin cement to Y-TZP after phosphoric acid pretreatment and different chemical conditioning protocols, and to test the durability of SBSs after long term storage and thermal cycling conditions. To investigate the effect of phosphoric acid 75 on the surface roughness of a yttrium-stabilized tetragonal zirconia polycrystal ceramic (Y-TZP). METHODS: Sixty bars of zirconia (30 X 5 X 2 mm) were polished and glued to acrylic tubes. Each bar received 3 different surface treatments (control, phosphoric acid and air-abrasion). The bars were randomly assigned into five groups according to different primers applied (No primer, Clearfil ceramic primer, Monobond Plus, Z-prime plus and AZ primer). After that, resin cement was mixed, injected into a mold, and light cured. The specimens were tested in shear on a universal testing machine. Half of the specimens were tested after 24 h water storage and the other half were tested after 90 days of water storage and thermocycling. Non-contact profilometer was used to assess the effect of the three surface pretreatments on surface roughness of zirconia. Mode of failure and surface roughness were evaluated with SEM. Statistical analyses were conducted with mixed-model ANOVA and chi-square tests. RESULTS: The results indicated that Monobond Plus primer showed higher SBS after phosphoric acid treatment compared to other primers used. SBS of all air abrasion groups were higher than phosphoric acid groups. Long term water storage and thermocycling led to a statistically significant decrease in SBS for all groups except Monobond Plus, Z-prime plus and AZ primer after air abrasion treatment. Surface roughness measurements showed no significant differences among the surface treatments. CONCLUSION: Monobond Plus provided the highest SBS to zirconia after phosphoric acid treatment. The findings suggest that the air abrasion is the best surface treatment after contamination of zirconia surface. Monobond Plus, Z-prime plus and AZ primer showed durable SBS after air abrasion treatment and aging process.

The purpose of the study was to get the essential information about the pre-primers that were taught in the Dallas County Schools, and to take a representative group and evaluate them as to the merits of the books for classroom use.

This thesis is based on the use of ISSR method for studying genetic diversity of oilseed rape. Altogether, 180 genotypes were evaluated. Frozen leaves of oilseed rape from three breeding station called: 1/ breeding station Selgen a.s., 2/ OSEVA PRO s.r.o. and 3/ breeding station Slapy u Tábora were used as imput material. DNA isolation was performed by Williams, who modified CTAB method for oilseed rape. We selected for ISSR 4 UBC which showed high variability and stability. The analysis was performed by gel electrophoresis. This thesis is also focused on comparison of gel electrophoresis and chip electrophoresis. In this thesis We show a matrix of simmilarity between varieties. The results were also provided back to breeders as a basis for further breeding.

abstract: Cooperativity can be used to manipulate binding affinities of DNA biosensors – improving specificity without sacrificing sensitivity; examples include tentacle probes (TPs) and cooperative primers (CPs). This thesis body of work: (1) used TPs to develop a rapid, low-cost diagnostic for detecting the point mutation leading to Navajo Neurohepatopathy (NNH) and (2) used CPs to amplify a symmetric bowtie-barcoded origami with captured t-cell receptor (TCR) α and β mRNA of a single cell. NNH (affecting 1-in-1600 Navajo babies) is a fatal genetic disorder often caused by 149G>A mutation and is characterized by brain damage and liver disease/failure. Phoenix Children’s Hospital currently uses gene sequencing to identify the 149G>A mutation. While this process is conclusive, there are limitations, as it requires both time (3-4 weeks) and money (>$700). Ultimately, these factors create barriers that can directly impact a patient’s quality of life. Assessment of the developed TP diagnostic, using genomic DNA derived from FFPE patient liver samples, suggests nearly 100% specificity and sensitivity while reducing cost to ~$250 (including cost of labor) and providing a diagnosis within 48 hours. TCR specificity is dependent on V(D)J recombination as well as pairing of the αβ chains. Drs. Schoettle and Blattman have developed a solution in which a bowtie-barcoded origami strand nanostructure is transfected into individual cells of a heterogeneous cell population to capture and protect αβ mRNA. When PCR of the origami template is performed with Vα, X, Vβ, and Y primers, the α and β gene segments cannot be tied back to a barcode – and paired. Assessment of the developed CPs for PCR suggests correct individual amplification using (1) Va + Xcp and (2) Vβ + Ycp primers, whereas combination of all the primers (Va, Xcp, Vb, and Ycp) suggests hybridization of the Vα + Xcp and Vβ + Ycp products due to the origami target symmetry. / Dissertation/Thesis / Third Iteration for Cooperative Primers / Masters Thesis Biomedical Engineering 2018

The present study is an analysis of six primers adopted for use in the public schools of Texas with the idea of comparing word-meanings used in the six books in order to find out possible sequences for the use of the books.

This study investigates the pheromone preference of a bark beetle predator Thanasimus dubius between latent and intermediate phases of southern pine beetle. Two trap treatments were set up in each SPB phase. Standard Lindgren funnel traps were baited with either SPB lures or Ips lures. The number of T. dubius caught in each trap was recorded and data was analyzed using both the Mann Whitney U test and a two-way factorial ANOVA. Thanasimus dubius showed no variation in SPB pheromone preference but did show a slight increase in preference for Ips pheromones in intermediate phase areas. A protocol was developed to identify prey DNA within gut contents of T. dubius to understand prey preference in relation to pheromone preference. Primers were developed to amplify CO1 gene sequences from five different bark beetles. All primers were specific to their own DNA and able to detect at least 0.2 picograms of DNA.

No description available.

A contaminação bacteriana por espécies dos gêneros Lactobacillus, Bacillus e Leuconostoc entre outras bactérias lácticas é um dos principais fatores que afetam o rendimento da fermentação alcoólica. A formação de biofilmes acaba protegendo as bactérias e é uma fonte permanente de contaminação. Objetivando caracterizar tais contaminações em (1) biofilmes de centrífuga, dorna, trocador de calor e tubulação de água e (2) melaço, mosto, levedo, levedo tratado (com H2SO4) e vinho, amostras foram coletadas em diferentes períodos de um sistema fermentativo de alto teor alcoólico (16%). As enzimas de restrição AluI, BstUI, HaeIII, HinfI, MseI e MspI utilizadas nas análises de T-RFLP foram definidas por análises in silico com sequências do gene 16S rRNA de contaminantes freqüentes. Essas enzimas geram uma maior quantidade de T-RFs únicos entre 30 e 650 pb. Os DNAs extraídos das amostras foram submetidos às análises de T-RFLP para obtenção do perfil molecular das comunidades microbianas dos pontos de coleta. Os índices de diversidade de Shannon foram calculados com base no número dos T-RFs. Foram realizadas as análises dos componentes principais (PCA) e a inferência filogenética dos contaminantes com base nos perfis dos T-RFs. A quantificação dos principais táxons contaminantes foi feita por qPCR utilizando primers específicos delineados neste estudo e considerando a média de cópias do gene 16S rRNA presentes no genoma de cada táxon bacteriano. Na primeira coleta o biofilme de água apresentou maior índice de diversidade microbiana e na segunda, melaço e mosto. PCA sugere que os biofilmes (e não as fontes externas) são os principais contaminantes desse processo fermentativo devido as suas semelhanças com a composição das outras comunidades analisadas. Espécies de Lactobacillus e Bacillus predominaram entre as amostras da primeira coleta. Halomonas, Streptococcus, Lactococcus e Pseudomonas foram detectados em amostras de biofilme e em amostras líquidas, sendo os principais contaminantes provindos de biofilme no momento da primeira coleta. Na segunda coleta Bacillus foi o principal contaminante e novamente gêneros produtores de ácido lático como Streptococcus, Lactobacillus e Staphylococcus foram os mais freqüentes. Os resultados concordam com o reportado na literatura sobre sistemas fermentativos convencionais. Apenas os primers desenhados para amplificação do gene 16S rRNA de Burkholderia, Pseudomonas e Weissella apresentaram especificidade em testes com isolados. Halomonas sp. foi encontrada em biofilme de dorna através do sequenciamento utilizando primers para esse gênero. Halomonas pode produzir levânio podendo haver o consumo da sacarose disponível para fermentação. Biofilme da centrífuga teve a maior quantidade de micro-organismos nos dois momentos de coleta (1,93E+06 UFC.mg-1 e 2,14E+07 UFC.mg-1, respectivamente) assim como as amostra de levedo entre as amostras líquidas (1,03E+08 UFC.ml-1 e 2,96E+06 UFC.ml-1, na primeira e segunda coleta, respectivamente), indicando níveis consideráveis de contaminantes. Burkholderia e Pseudomonas foram os mais abundantes entre as amostras de biofilmes da primeira e segunda coleta. Nas amostras líquidas Burkholderia apresentou-se em maior quantidade na maioria das amostras da primeira coleta; enquanto Pseudomonas e Weissella em geral predominaram equivalentemente entre as amostras da segunda coleta / Bacterial contamination by Lactobacillus, Bacillus and Leuconostoc and other lactic acid bacteria is one of the main factors that affects the yield in alcoholic fermentation process. Biofilm formation protects the bacteria community and it is a permanent source of contamination. For characterization of these contaminations in (1) biofilms from centrifuge, tank fermentation, heat exchanger and water pipe and (2) molasses, must, yeast, yeast treated (with H2SO4) and wine, samples were taken at two different periods from fermentation system characterized by high alcohol yields (16%). Restriction enzymes AluI, BstUI, HaeIII, HinfI, MseI and MspI used in T-RFLP analysis were defined by 16S rRNA gene sequences analysis in silico from common contaminants. These enzymes generate high number of unique T-RFs between 30 and 650 bp. DNA from samples were used as template in T-RFLP reactions in order to obtain molecular profiles of microbial communities present at each sample. Shannon diversity index was calculated based on T-RFs numbers. Principal component analysis (PCA) and phylogenetic inference of contaminants were performed based on T-RFs profiles. The main contaminant bacterial taxa were quantified by qPCR using specific primers designed in this study and considering the average of 16S rRNA gene copies previously counted into the genome of each bacterial taxon. Water pipe biofilm showed the highest rate of bacterial diversity in the samples collected in the first sampling period. For the samples collected in the second sampling, the highest rate of bacterial diversity was revealed for molasses and must. PCA suggested that biofilms (but not external sources) are the main contaminants in the studied fermentation process. It is probably due their similarities with the composition of other analyzed communities. Lactobacillus and Bacillus species predominated in first sampling period. Halomonas, Streptococcus, Lactococcus and Pseudomonas were detected in biofilm and liquid samples. They were the main contaminants from biofilm at this time of sampling. In the second sampling period, Bacillus was the most common genera and other lactic acid bacteria such Streptococcus, Staphylococcus and Lactobacillus were also the most frequent contaminants. These results agree with other reported in the literature about conventional fermentation systems. Only the primers designed in this study to amplify the 16S rRNA gene of Burkholderia, Pseudomonas and Weissella showed specificity in tests with bacterial strains. Halomonas sp. was revealed in biofilms from tank fermentation by DNA sequencing using designed primers for genera. Halomonas can produce levan and may consume sucrose available for generation of alcohol. Centrifugal biofilm had the highest amount of bacteria in both sampling periods (1.93E+06 CFU.mg-1 and 2.14E+07 CFU.mg-1, respectively). In liquid samples, yeast had the highest amount of bacteria in both sampling periods (1.03E+08 CFU.ml-1 and 2.96E+06 CFU.ml-1, respectively); it shows significant levels of contaminants. Burkholderia and Pseudomonas were more abundant among biofilm samples of all samplings. Burkholderia was present in high quantities in the majority of liquid samples taken during the first sampling period; Pseudomonas and Weissella equivalently predominated among samples taken during the second sampling period